Indole sPLA2 inhibitors

ABSTRACT

A class of novel acylsulfonamide substituted indole compounds is disclosed together with the use of such compounds for inhibiting sPLA 2  mediated release of fatty acids for treatment of inflammatory diseases such as septic shock.

This application is a divisional of U.S. patent application Ser. No.09/762,070 filed 30 Jan. 2001, now U.S. Pat. No. 6,608,099, which is a371 of PCT/US99/17460 filed 2 Aug. 1999 which claims the benefit of U.S.Provisional Patent Application 60/095,109 filed 3 Aug. 1998.

FIELD OF THE INVENTION

This invention relates to novel indole compounds useful for inflammatorydiseases.

BACKGROUND OF THE INVENTION

The structure and physical properties of human non-pancreatic secretoryphospholipase A₂ (hereinafter called, “sPLA₂”) has been thoroughlydescribed in two articles, namely, “Cloning and Recombinant Expressionof Phospholipase A₂ Present in Rheumatoid Arthritic Synovial Fluid” bySeilhamer, Jeffrey J.; Pruzanski, Waldemar; Vadas Peter; Plant, Shelley;Miller, Judy A.; Kloss, Jean; and Johnson, Lorin K.; The Journal ofBiological Chemistry, Vol. 264, No. 10, Issue of April 5, pp. 5335-5338,1989; and “Structure and Properties of a Human Non-pancreaticPhospholipase A₂” by Kramer, Ruth M.; Hession, Catherine; Johansen,Berit; Hayes, Gretchen; McGray, Paula; Chow, E. Pingchang; Tizard,Richard; and Pepinsky, R. Blake; The Journal of Biological Chemistry,Vol. 264, No. 10, Issue of April 5, pp. 5768-5775, 1989; the disclosuresof which are incorporated herein by reference.

Indole type sPLA2 inhibitors having gyloxylamide, acetamide andhydrazide substituents are described in U.S. Pat. Nos. 5,654,326;5,684,034; and 5,578,634 respectively.

It is believed that sPLA₂ is a rate limiting enzyme in the arachidonicacid cascade which hydrolyzes membrane phospholipids. Thus, it isimportant to develop compounds which inhibit sPLA₂ mediated release offatty acids (e.g., arachidonic acid). Such compounds would be of valuein general treatment of conditions induced and/or maintained byoverproduction of sPLA₂; such as sepsis or rheumatoid arthritis.

It is desirable to develop new compounds and treatments for sPLA₂induced diseases.

SUMMARY OF THE INVENTION

This invention is a class of novel indole compounds to inhibit mammaliansPLA₂ mediated release of fatty acids.

This invention is also a novel class of indole compounds having potentand selective effectiveness as inhibitors of mammalian sPLA₂.

This invention is also a indole compound useful in the treatment ofInflammatory Diseases.

This invention is also a pharmaceutical composition containing novelindoles of the invention.

This invention is also a method of preventing and treating InflammatoryDiseases in mammals by administration of a therapeutically effectiveamount of the indole of the invention.

This invention is also the indole compounds of the invention orcompositions comprising the compounds of the invention as activeingredient for use as a medicament in the treatment of InflammatoryDiseases.

Definitions:

The term, “Inflammatory Diseases” refers to diseases such asinflammatory bowel disease, sepsis, septic shock, adult respiratorydistress syndrome, pancreatitis, trauma-induced shock, bronchial asthma,allergic rhinitis, rheumatoid arthritis, cystic fibrosis, stroke, acutebronchitis, chronic bronchitis, acute bronchiolitis, chronicbronchiolitis, osteoarthritis, gout, spondylarthropathris, ankylosingspondylitis, Reiter's syndrome, psoriatic arthropathy, enterapathricspondylitis, Juvenile arthropathy or juvenile ankylosing spondylitis,Reactive arthropathy, infectious or post-infectious arthritis,gonoccocal arthritis, tuberculous arthritis, viral arthritis, fungalarthritis, syphilitic arthritis, Lyme disease, arthritis associated with“vasculitic syndromes”, polyarteritis nodosa, hypersensitivityvasculitis, Luegenec's granulomatosis, polymyalgin rheumatica, jointcell arteritis, calcium crystal deposition arthropathris, pseudo gout,non-articular rheumatism, bursitis, tenosynomitis, epicondylitis (tenniselbow), carpal tunnel syndrome, repetitive use injury (typing),miscellaneous forms of arthritis, neuropathic joint disease (charco andjoint), hemarthrosis (hemarthrosic), Henoch-Schonlein Purpura,hypertrophic osteoarthropathy, multicentric reticulohistiocytosis,arthritis associated with certain diseases, surcoilosis,hemochromatosis, sickle cell disease and other hemoglobinopathries,hyperlipoproteineimia, hypogammaglobulinemia, hyperparathyroidism,acromegaly, familial Mediterranean fever, Behat's Disease, systemiclupus erythrematosis, or relapsing polychondritis and related diseaseswhich comprises administering to a mammal in need of such treatment atherapeutically effective amount of the compound of formula I in anamount sufficient to inhibit sPLA₂ mediated release of fatty acid and tothereby inhibit or prevent the arachidonic acid cascade and itsdeleterious products.

The term, “indole nucleus” refers to a nucleus (having numberedpositions)with the structural formula (X):

The indole compounds of the invention employ certain defining terms asfollows:

The term, “alkyl” by itself or as part of another substituent means,unless otherwise defined, a straight or branched chain monovalenthydrocarbon radical such as methyl, ethyl, n-propyl, isopropyl, n-butyl,tertiary butyl, sec-butyl, n-pentyl, and n-hexyl.

The term, “alkenyl” employed alone or in combination with other termsmeans a straight chain or branched monovalent hydrocarbon group havingthe stated number range of carbon atoms, and typified by groups such asvinyl, propenyl, crotonyl, isopentenyl, and various butenyl isomers.

The term, “halo” means fluoro, chloro, bromo, or iodo. The term,heterocyclic radical, refers to radicals derived from monocyclic orpolycyclic, saturated or unsaturated, substituted or unsubstitutedheterocyclic nuclei having 5 to 14 ring atoms and containing from 1 to 3hetero atoms selected from the group consisting of nitrogen, oxygen orsulfur. Typical heterocyclic radicals are pyrrolyl, pyrrolodinyl,piperidinyl, furanyl, thiophenyl, pyrazolyl, imidazolyl,phenylimidazolyl, triazolyl, isoxazolyl, oxazolyl, thiazolyl,thiadiazolyl, indolyl, carbazolyl, norharmanyl, azaindolyl,benzofuranyl, dibenzofuranyl, dibenzothiophenyl, indazolyl,imidazo(1.2-A)pyridinyl, benzotriazolyl, anthranilyl,1,2-benzisoxazolyl, benzoxazolyl, benzothiazolyl, purinyl, pyridinyl,dipyridylyl, phenylpyridinyl, benzylpyridinyl, pyrimidinyl,phenylpyrimidinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl, phthalazinyl,quinazolinyl,morpholino, thiomorpholino, homopiperazinyl,tetrahydrofuranyl, tetrahydropyranyl, oxacanyl, 1,3-dioxolanyl,1,3-dioxanyl, 1,4-dioxanyl, tetrahydrothiopheneyl,pentamethylenesulfadyl, 1,3-dithianyl, 1,4-dithianyl, 1,4-thioxanyl,azetidinyl, hexamethyleneiminium, heptamethyleneiminium, piperazinyl andquinoxalinyl.

The term, “carbocyclic radical” refers to radicals derived from asaturated or unsaturated, substituted or unsubstituted 5 to 14 memberedorganic nucleus whose ring forming atoms (other than hydrogen) aresolely carbon atoms. Typical carbocyclic radicals are cycloalkyl,cycloalkenyl, phenyl, spiro[5.5]undecanyl, naphthyl, norbornanyl,bicycloheptadienyl, tolulyl, xylenyl, indenyl, stilbenyl, terphenylyl,diphenylethylenyl, phenyl-cyclohexenyl, acenaphthylenyl, andanthracenyl, biphenyl, bibenzylyl and related bibenzylyl homologuesrepresented by the formula (a):

where n is a number from 1 to 8.

The term, “non-interfering substituent”, refers to radicals suitable forsubstitution at positions 4,5,6 and/or 7 of the indole nucleus and onother nucleus substituents (as hereinafter described for Formula I), andradicals suitable for substitution on the heterocyclic radical andcarbocyclic radical as defined above. Illustrative non-interferingradicals are C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₇-C₁₂ aralkyl,C₇-C₁₂ alkaryl, C₃-C₈ cycloalkyl, C₃-C₈ cycloalkenyl, phenyl, tolulyl,xylenyl, biphenyl, C₁-C₈ alkoxy, C₂-C₈ alkenyloxy, C₂-C₈ alkynyloxy,C₂-C₁₂ alkoxyalkyl, C₂-C₁₂ alkoxyalkyloxy, C₂-C₁₂ alkylcarbonyl, C₂-C₁₂alkylcarbonylamino, C₂-C₁₂ alkoxyamino, C₂-C₁₂ alkoxyaminocarbonyl,C₁-C₁₂ alkylamino, C₁-C₆ alkylthio, C₂-C₁₂ alkylthiocarbonyl, C₁-C₈alkylsulfinyl, C₁-C₈ alkylsulfonyl, C₂-C₈ haloalkoxy, C₁-C₈haloalkylsulfonyl, C₂-C₈ haloalkyl, C₁-C₈ hydroxyalkyl, —C(O)O(C₁-CBalkyl), —(CH₂)_(n)—O—(C₁-C₈ alkyl), benzyloxy, phenoxy, phenylthio,—(CONHSO₂R), —CHO, amino, amidino, bromo, carbamyl, carboxyl,carbalkoxy, —(CH₂)_(n)—CO₂H, chloro, cyano, cyanoguanidinyl, fluoro,guanidino, hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo,nitro, phosphono, —SO₃H, thioacetal, thiocarbonyl, and C₁-C₈ carbonyl;where R is C₁-C₈ alkyl and n is from 1 to 8.

The term “substituted group” is an organic group substituted with one ormore non-interfering substituents.

The term, “(acylsulfonamide group)” is a group represented by theformula:

where R₈₁ is —CF₃ or an organic substituent consisting of carbon andhydrogen with or without oxygen, nitrogen, sulfur, halogen or otherelements.

The term “indole acylsulfonamide compound” is synonymous with the term“indole compound.” The words, “linker” refer to a divalent linking groupsymbolized as, -(L_(a))-, which has the function of joining the 4 or 5position of the indole nucleus to an acylsulfonamide group in thegeneral relationship:

The words, “linker length”, refer to the number of atoms (excludinghydrogen) in the shortest chain of the linking group -(L_(a))- thatconnects the 4 or 5 position of the indole nucleus with theacylsulfonamide group. The presence of a carbocyclic ring in -(L_(a))-counts as the number of atoms approximately equivalent to the calculateddiameter of the carbocyclic ring. Thus, a benzene or cyclohexane ring inthe linker counts as 2 atoms in calculating the length of -(L_(a))-.Illustrative linker groups are;

wherein, groups (a), (b), and (c) have linker lengths of 5, 7, and 2,respectively.

The term, “amine”, includes primary, secondary and tertiary amines.

The terms, “mammal” and “mammalian” include human.

The term, “alkylene chain of 1 or 2 carbon atoms” refers to the divalentradicals, —CH₂—CH₂— and —CH₂—.

The term, “group containing 1 to 4 non-hydrogen atoms” refers torelatively small groups which form substituents at the 2 position of theindole nucleus, said groups may contain non-hydrogen atoms alone, ornon-hydrogen atoms plus hydrogen atoms as required to satisfy theunsubstituted valence of the non-hydrogen atoms, for example; (i) groupsabsent hydrogen which contain no more than 4 non-hydrogen atoms such as—CF₃, —Cl, —Br, —NO₂, —CN, —SO₃; and (ii) groups having hydrogen atomswhich contain less than 4 non-hydrogen atoms such as —CH₃, —C₂H₅, and—CH═CH₂.

The term “spiro[5.5]undecanyl” refers to the group represented by theformula;

The term “organic substituent” is a monovalent radical consisting ofcarbon and hydrogen with or without oxygen, nitrogen, sulfur, halogen orother element. Illustrative organic substituents are —CH₃, —C₂H₅,—CH₂OCH₃, and —CH₂SCH₃.

The Indole Compounds of the Invention:

The compounds of the invention have the general formula (I) or apharmaceutically acceptable salt or solvate thereof;

wherein;

each X is independently oxygen or sulfur;

R₁ is selected from groups (a), (b) and (c) wherein;

-   -   (a) is C₇-C₂₀ alkyl, C₇-C₂₀ haloalkyl, C₇-C₂₀ alkenyl, C₇-C₂₀        alkynyl, carbocyclic radical, or heterocyclic radical, or    -   (b) is a member of (a) substituted with one or more        independently selected non-interfering substituents; or    -   (c) is the group -(L₁)—R₁₁; where, -(L₁)- is a divalent linking        group of 1 to 8 atoms and where R₁₁ is a group selected from (a)        or (b);

R₂ is hydrogen, or a group containing 1 to 4 non-hydrogen atoms plus anyrequired hydrogen atoms;

R₃ is -(L₃)-Z, where -(L₃)- is a divalent linker group selected from abond or a divalent group selected from:

and Z is selected from acetamide, thioacetamide, glyoxylamide,thioglyoxylamide, hydrazide or thiohydrazide groups represented by theformulae,

where R₃₁ and R₃₂ are independently selected from hydrogen, C₁-C₈ alkyl,C₁-C₈ haloalkyl, and C₃-C₄ cycloalkyl, and X is oxygen or sulfur;

R₄ and R₅ are independently selected from hydrogen, a non-interferingsubstituent, or the group, -(L_(a))-(acylsulfonamide group); wherein-(L_(a))-, is a linker having a linker length of 1 to 8, provided, thatat least one of R₄ and R₅ must be the group, -(L_(a))-(acylsulfonamidegroup); and the (acylsulfonamide group) is represented by the formula;

where R₈₁ is selected from the group consisting of C₁-C₈ alkyl, C₁-C₈alkoxy, C₁-C₈ alkylthio, C₁-C₈ alkylamino, C₁-C₈ haloalkyl, C₁-C₁₄aralkyl, C₁-C₁₄ alkylaryl, aryl, thioaryl, C₃-C₁₄ carbocyclic radical,C₃-C₁₄ heterocyclic radical.

R₆ and R₇ are selected from hydrogen, non-interfering substituent,carbocyclic radical, carbocyclic radical substituted withnon-interfering substituent(s), heterocyclic radical, and heterocyclicradical substituted with non-interfering substituent(s).

Preferred Subgroups of Compounds of Formula (I):

I. Preferred R₁ Substituents:

A preferred subclass of compounds of formula (I) wherein for R₁ thedivalent linking group -(L₁)- are those corresponding to the formulae(VIIa), (VIIb), (VIIc), (VIId), (VIIe), and (VIIf):

where Q₁ is a bond or any of the divalent groups (VIIa), (VIIb), (VIIc),(VIId), (VIIe), and (VIIf) and each R₁₀ is independently hydrogen, C₁₋₈alkyl, C₁₋₈ haloalkyl or C₁₋₈ alkoxy.

Particularly preferred as the linking group -(L₁)- of R₁ is an alkylenechain of 1 or 2 carbon atoms, namely, —(CH₂)— or —(CH₂—CH₂)—.

The preferred group for R₁₁ is a substituted or unsubstituted groupselected from the group consisting of C₅-C₁₄ cycloalkyl, C₅-C₁₄cycloalkenyl, phenyl, naphthyl, norbornanyl, bicycloheptadienyl,tolulyl, xylenyl, indenyl, stilbenyl, terphenylyl, diphenylethylenyl,phenyl-cyclohexenyl, acenaphthylenyl, and anthracenyl, biphenyl,bibenzylyl and related bibenzylyl homologues represented by the formula(a);

where n is a number from 1 to 8.

Particularly preferred are compounds wherein for R₁ the combined group-(L₁)—R₁₁ is selected from the group consisting of

where R₁₂ is a radical independently selected from halo, C₁-C₈ alkyl,C₁-C₈ alkoxy, —S—(C₁-C₈ alkyl), —O—(C₁-C₈ alkyl) and C₁-C₈ haloalkylwhere t is a number from 0 to 5 and u is a number from 0 to 4.II. Preferred R₂ Substituents:

R₂ is preferably selected from the group consisting of hydrogen, C₁-C₄alkyl, C₂-C₄ alkenyl, —O—(C₁-C₃ alkyl),

—S—(C₁-C₃ alkyl), —C₁-C₄ cycloalkyl —CF₃, halo, —NO₂, —CN, —SO₃.Particularly preferred R₂ groups are selected from hydrogen, methyl,ethyl, propyl, isopropyl, cyclopropyl, —F, —CF₃, —Cl, —Br, or —O—CH₃.

III. Preferred R₃ Substituents:

A preferred subclass of compounds of formula (I) are those wherein X isoxygen.

Another preferred subclass of compounds of formula (I) are those whereinZ is a glyoxylamide group.

Most preferred are compounds of formula (I) wherein R₃ is the glyoxamidegroup. For the group R₃ it is preferred that the linking group -(L₃)- bea bond.IV. Preferred R₄ Substituents:

Another preferred subclass of compounds of formula (I) are those whereinR₄ is a substituent having a linker with a linker length of 2 or 3 andthe linker group, -(L₄)-, for R₄ is selected from a group represented bythe formula;

where Q₂ is selected from the group —(CH₂)—, —O—, —NH—, —C(O)—, and —S—,and each R₄₀ is independently selected from hydrogen, C₁-C₈ alkyl, aryl,C₁-C₈ alkaryl, C₁-C₈ alkoxy, aralkyl, and halo. Most preferred arecompounds where the linker, -(L₄)-, for R₄ is selected from the specificgroups;

where R₄₀ is hydrogen or C₁-C₈ alkyl.

Preferred as the (acylsulfonamide group) in the group R₄ areacylsulfonamide groups selected from:

where R₈₁ is selected from —CF₃, C₁-C₈ alkyl, C₁-C₈ alkoxy, C₁-C₈alkylthio, C₁-C₈ alkylamino, C₁-C₈ haloalkyl, C₁-C₁₄ aralkyl, C₁-C₁₄alkylaryl, aryl, thioaryl, C₃-C₁₄ carbocyclic radical, C₃-C₁₄heterocyclic radical. A salt or a solvate derivative of the(acylsulfonamide group) is also suitable.

Particularly preferred are acylsulfonamide groups selected from:

or salts or solvates thereof.V. Preferred R₅ Substituents:

Preferred linker, -(L_(a))-, for R₅ is selected from the groupconsisting of;

wherein R₅₄, R₅₅, R₅₆ and R₅₇ are each independently hydrogen, C₁-C₈alkyl, C₁-C₈ haloalkyl, aryl, C₁-C₈ alkoxy, or halo.VI. Preferred (Acylsulfonamide Group) for R₄ and/or R₅ Substitutions:

At least one of R₄ and R₅ must be the group, -(L_(a))-(acylsulfonamidegroup). The preferred (acylsulfonamide group) on the group-(L_(a))-(acylsulfonamide group) of R₄ or R₅ is selected from

In addition, it is preferred that only one R₄ or R₅ substituents be thegroup, -(L_(a))-(acylsulfonamide group).Most preferred is that the R₄ substituent be the group,-(L_(a))-(acylsulfonamide group).V. Preferred R₆ Substituents:

Another preferred subclass of compounds of formula (I) are those whereinfor R₆ the non-interfering substituent is methyl, ethyl, propyl,isopropyl, —S—CH₃, —O—CH₃, C₄-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₇-C₁₂ aralkyl, C₇-C₁₂ alkaryl, C₃-C₈ cycloalkyl, C₃-C₈ cycloalkenyl,phenyl, tolulyl, xylenyl, biphenyl, C₁-C₆ alkoxy, C₂-C₆ alkenyloxy,C₂-C₆ alkynyloxy, C₂-C₁₂ alkoxyalkyl, C₂-C₁₂ alkoxyalkyloxy, C₂-C₁₂alkylcarbonyl, C₂-C₁₂ alkylcarbonylamino, C₂-C₁₂ alkoxyamino, C₂-C₁₂alkoxyaminocarbonyl, C₁-C₁₂ alkylamino, C₁-C₆ alkylthio, C₂-C₁₂alkylthiocarbonyl, C₁-C₆ alkylsulfinyl, C₁-C₆ alkylsulfonyl, C₂-C₆haloalkoxy, C₁-C₆ haloalkylsulfonyl, C₂-C₆ haloalkyl, C₁-C₆hydroxyalkyl, —C(O)O(C₁-C₆ alkyl), —(CH₂)_(n)—O—(C₁-C₆ alkyl),benzyloxy, phenoxy, phenylthio, —(CONHSO₂R), —CHO, amino, amidino,bromo, carbamyl, carboxyl, carbalkoxy, —(CH₂)_(n)—CO₂H, chloro, cyano,cyanoguanidinyl, fluoro, guanidino, hydrazide, hydrazino, hydrazido,hydroxy, hydroxyamino, iodo, nitro, phosphono, —SO₃H, thioacetal,thiocarbonyl, and C₁-C₆ carbonyl; where R is C₁-C₈ alkyl and n is from 1to 8.

Preferred compounds of the invention are those having the generalformula (II), or a pharmaceutically acceptable salt or solvatederivative thereof;

wherein;

R₈₁ is selected from C₁-C₈ alkyl, C₁-C₈ alkoxy, C₁-C₈ alkylthio C₁-C₈haloalkyl, C₁-C₈ alkylamino, C₁-C₁₄ aralkyl, C₁-C₁₄ alkylaryl, aryl,thioaryl, C₃-C₁₄ carbocycle, C₃-C₁₄ heterocycle.

R₂₂ is selected from hydrogen, methyl, ethyl, propyl, isopropyl,cyclopropyl, —F, —CF₃, —Cl, —Br, or —O—CH₃;

-(L₄)- is a divalent group selected from;

where R₄₀, R₄₁, R₄₂, and R₄₃ are each independently selected fromhydrogen or C₁-C₈ alkyl.

R₁₆ is selected from hydrogen, C₁-C₈ alkyl, C₁-C₈ alkoxy, C₁-C₈alkylthio C₁-C₈ haloalkyl, C₁-C₈ hydroxyalkyl, and halo.

R₁₃ is selected from hydrogen and C₁-C₈ alkyl, C₁-C₈ alkoxy, —S—(C₁-C₈alkyl), C₁-C₈ haloalkyl, C₁-C₈ hydroxyalkyl, and halo, and t is aninteger from 0 to 5.

A preferred compound (and all pharmaceutically acceptable salts andsolvate derivatives thereof) which is illustrative of the compounds ofthe invention is as follows:

The salts of the above indole compounds represented by formulae (I) and(II) are an additional aspect of the invention. In those instances wherethe compounds of the invention possess acidic or basic functional groupsvarious salts may be formed which are more water soluble andphysiologically suitable than the parent compound. Representativepharmaceutically acceptable salts, include but are not limited to, thealkali and alkaline earth salts such as lithium, sodium, potassium,calcium, magnesium, aluminum and the like. Salts are convenientlyprepared from the free acid by treating the acylsulfonamide in solutionwith a base or by exposing the acylsulfonamide to an ion exchange resinFor example, the (acylsulfonamide group) of the substituent R₄ ofFormula I may be selected as —(CO)NHSO₂CH₃ and salts may be formed byreaction with appropriate bases (e.g., NaOH, KOH) to yield thecorresponding sodium and potassium salt.

Included within the definition of pharmaceutically acceptable salts arethe relatively non-toxic, inorganic and organic base addition salts ofcompounds of the present invention, for example, ammonium, quaternaryammonium, and amine cations, derived from nitrogenous bases ofsufficient basicity to form salts with the compounds of this invention(see, for example, S. M. Berge, et al., “Pharmaceutical Salts,” J. Phar.Sci., 66: 1-19 (1977)). Moreover, the basic group(s) of the compound ofthe invention may be reacted with suitable organic or inorganic acids toform salts such as acetate, benzenesulfonate, benzoate, bicarbonate,bisulfate, bitartrate, borate, bromide, camsylate, carbonate, chloride,clavulanate, citrate, chloride, edetate, edisylate, estolate, esylate,fluoride, fumarate, gluceptate, gluconate, glutamate,glycolylarsanilate, hexylresorcinate, bromide, chloride,hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate,malate, malseate, mandelate, mesylate, methylbromide, methylnitrate,methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, palmitate,pantothenate, phosphate, polygalacturonate, salicylate, stearate,subacetate, succinate, tannate, tartrate, tosylate, trifluoroacetate,trifluoromethane sulfonate, and valerate.

Certain compounds of the invention may possess one or more chiralcenters and may thus exist in optically active forms. Likewise, when thecompounds contain an alkenyl or alkenylene group there exists thepossibility of cis- and trans-isomeric forms of the compounds. The R—and S— isomers and mixtures thereof, including racemic mixtures as wellas mixtures of cis- and trans-isomers, are contemplated by thisinvention. Additional asymmetric carbon atoms can be present in asubstituent group such as an alkyl group. All such isomers as well asthe mixtures thereof are intended to be included in the invention. If aparticular stereoisomer is desired, it can be prepared by methods wellknown in the art by using stereospecific reactions with startingmaterials which contain the asymmetric centers and are already resolvedor, alternatively by methods which lead to mixtures of the stereoisomersand subsequent resolution by known methods. For example, a racemicmixture may be reacted with a single enantiomer of some other compound.This changes the racemic form into a mixture of diastereomers anddiastereomers, because they have different melting points, differentboiling points, and different solubilities can be separated byconventional means, such as crystallization.

Method of Making the Compounds of the Invention:

The synthesis of the indole compound of the invention (viz., Compoundsof Formulae I and II) can be accomplished by well known methods asrecorded in the chemical literature. In particular, the indole startingmaterials may be prepared by the synthesis schemes taught in U.S. Pat.No. 5,654,326; the disclosure of which is incorporated herein byreference. Procedures useful for the synthesis of the starting materialare shown in the Scheme below:

wherein, for the preceding Scheme, R₆ is an organic substituent or —CF₃,R₄ is equivalent to R₆ or R₇ in Formula I, R₂ is equivalent to R₂ informula I, R₃ is equivalent to (R₁₂)_(t) in Formula I and R₅ is —H orC₁-C₈ alkyl.

To obtain the glyoxylamides substituted in the 4-position with an acidicfunction through an oxygen atom, the reactions outlined in scheme 1 areused (for conversions 1 thru 5, see ref. Robin D. Clark, Joseph M.Muchowski, Lawrence E. Fisher, Lee A. Flippin, David B. Repke, MichelSouchet, Synthesis, 1991, 871-878, the disclosures of which areincorporated herein by reference). The ortho-nitrotoluene, 1, is readilyreduced to the 2-methylaniline, 2, using Pd/C as catalyst. The reductioncan be carried out in ethanol or tetrahydrofuran (THF) or a combinationof both, using a low pressure of hydrogen. The aniline, 2, on heatingwith di-tert-butyl dicarbonate in THF at reflux temperature is convertedto the N-tert-butylcarbonyl derivative, 3, in good yield. The dilithiumsalt of the dianion of 3 is generated at −40 to −20° C. in THF usingsec-butyl lithium and reacted with the appropriately substitutedN-methoxy-N-methylalkanamide. This product, 4, may be purified bycrystallization from hexane, or reacted directly with trifluoroaceticacid in methylene chloride to give the 1,3-unsubstituted indole 5. The1,3-unsubstituted indole 5 is reacted with sodium hydride indimethylformamide at room temperature (20-25° C.) for 0.5-1.0 hour. Theresulting sodium salt of 5 is treated with an equivalent of arylmethylhalide and the mixture stirred at a temperature range of 0-100° C.,usually at ambient room temperature, for a period of 4 to 36 hours togive the 1-arylmethylindole, 6. This indole, 6, is O-demethylated bystirring with boron tribromide in methylene chloride for approximately 5hours (see ref. Tsung-Ying Shem and Charles A Winter, Adv. Drug Res.,1977, 12, 176, the disclosure of which is incorporated herein byreference). The 4-hydroxyindole, 7, is alkylated with an alphabromoalkanoic acid ester in dimethylformamide (DMF) using sodium hydrideas a base, with reactions conditions similar to that described for theconversion of 5 to 6. The α-[(indol-4-yl)oxy]alkanoic acid ester, 8, isreacted with oxalyl chloride in methylene chloride to give 9, which isnot purified but reacted directly with ammonia to give the glyoxamide10. This product is hydrolyzed using 1N sodium hydroxide in MeOH. Theglyoxylamide, 11, is isolated either as the free carboxylic acid.Sulfonamide formation is accomplished by reaction of the appropriatesulfonamide with the indole acid (glyoxylamide) 11, in the presence of asuitable coupling agent such as EDCI(1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) anddimethylaminopyridine as a base to afford the indole acylsulfonamide,12. The formation of the sulfonamide, 12, can be accomplished by otherprocedures known in the art.

An alternative method of making starting materials substituted in the4-position is described in provisional patent application No. 60/082,110filed Apr. 17, 1998 and entitled “PROCESS FOR PREPARING4-SUBSTITUTED-1H-INDOLE-3-GLYOXYLAMIDES.” The esters described thereinmay be hydrolyzed to the acid form suitable for a starting material forpreparing compounds of the invention.

Methods of Using the Compounds of the Invention:

The method of the invention for inhibiting sPLA₂ mediated release offatty acids comprises contacting mammalian sPLA₂ with an therapeuticallyeffective amount of indole compounds corresponding to Formulae (I) or(II) as described herein including salt or a prodrug derivative thereof.

Another aspect of this invention is a method for treating InflammatoryDiseases such as inflammatory bowel disease, septic shock, adultrespiratory distress syndrome, pancreatitis, trauma, bronchial asthma,allergic rhinitis, rheumatoid arthritis, osteoarthritis, and relateddiseases which comprises administering to a mammal. (including a human)a therapeutically effective dose of the indole compound of the invention(see, formula I and II).

As previously noted the compounds of this invention are useful forinhibiting sPLA₂ mediated release of fatty acids such as arachidonicacid. By the term, “inhibiting” is meant the prevention ortherapeutically significant reduction in release of sPLA₂ initiatedfatty acids by the compounds of the invention. By “pharmaceuticallyacceptable” it is meant the carrier, diluent or excipient must becompatible with the other ingredients of the formulation and notdeleterious to the recipient thereof.

The specific dose of a compound administered according to this inventionto obtain therapeutic or prophylactic effects will, of course, bedetermined by the particular circumstances surrounding the case,including, for example, the compound administered, the route ofadministration and the condition being treated. Typical daily doses willcontain a non-toxic dosage level of from about 0.01 mg/kg to about 50mg/kg of body weight of an active compound of this invention.

Preferably compounds of the invention (per Formulae I or II) orpharmaceutical formulations containing these compounds are in unitdosage form for administration to a mammal. The unit dosage form can bea capsule or tablet itself, or the appropriate number of any of these.The quantity of Active ingredient in a unit dose of composition may bevaried or adjusted from about 0.1 to about 1000 milligrams or moreaccording to the particular treatment involved. It may be appreciatedthat it may be necessary to make routine variations to the dosagedepending on the age and condition of the patient. The dosage will alsodepend on the route of administration.

The compound can be administered by a variety of routes including oral,aerosol, rectal, transdermal, subcutaneous, intravenous, intramuscular,and intranasal.

Pharmaceutical formulations of the invention are prepared by combining(e.g., mixing) a therapeutically effective amount of the indole compoundof the invention together with a pharmaceutically acceptable carrier ordiluent therefor. The present pharmaceutical formulations are preparedby known procedures using well known and readily available ingredients.

In making the compositions of the present invention, the Activeingredient will usually be admixed with a carrier, or diluted by acarrier, or enclosed within a carrier which may be in the form of acapsule, sachet, paper or other container. When the carrier serves as adiluent, it may be a solid, semi-solid or liquid material which acts asa vehicle, or can be in the form of tablets, pills, powders, lozenges,elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solidor in a liquid medium), or ointment, containing, for example, up to 10%by weight of the active compound. The compounds of the present inventionare preferably formulated prior to administration.

For the pharmaceutical formulations any suitable carrier known in theart can be used. In such a formulation, the carrier may be a solid,liquid, or mixture of a solid and a liquid. For example, for intravenousinjection the compounds of the invention may be dissolved in at aconcentration of 2 mg/ml in a 4% dextrose/0.5% Na citrate aqueoussolution. Solid form formulations include powders, tablets and capsules.A solid carrier can be one or more substances which may also act asflavoring agents, lubricants, solubilisers, suspending agents, binders,tablet disintegrating agents and encapsulating material.

Tablets for oral administration may contain suitable excipients such ascalcium carbonate, sodium carbonate, lactose, calcium phosphate,together with disintegrating agents, such as maize, starch, or alginicacid, and/or binding agents, for example, gelatin or acacia, andlubricating agents such as magnesium stearate, stearic acid, or talc.

In powders the carrier is a finely divided solid which is in admixturewith the finely divided Active ingredient. In tablets the Activeingredient is mixed with a carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired. The powders and tablets preferably contain from about 1 toabout 99 weight percent of the Active ingredient which is the novelcompound of this invention. Suitable solid carriers are magnesiumcarbonate, magnesium stearate, talc, sugar lactose, pectin, dextrin,starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethylcellulose, low melting waxes, and cocoa butter.

Sterile liquid form formulations include suspensions, emulsions, syrupsand elixirs.

The Active ingredient can be dissolved or suspended in apharmaceutically acceptable carrier, such as sterile water, sterileorganic solvent or a mixture of both. The Active ingredient can often bedissolved in a suitable organic solvent, for instance aqueous propyleneglycol. Other compositions can be made by dispersing the finely dividedActive ingredient in aqueous starch or sodium carboxymethyl cellulosesolution or in a suitable oil.

The following pharmaceutical formulations 1 thru 8 are illustrative onlyand are not intended to limit the scope of the invention in any way.“Active ingredient”, refers to a compound according to Formula (I) or(II) or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

Quantity (mg/capsule) Formulation 1 Hard gelatin capsules are preparedusing the following ingredients: Active ingredient 250 Starch, dried 200Magnesium stearate 10 Total 460 mg Formulation 2 A tablet is preparedusing the ingredients below: Active ingredient 250 Cellulose,microcrystalline 400 Silicon dioxide, fumed 10 Stearic acid 5 Total 665mgThe components are blended and compressed to form tablets each weighing665 mg

Formulation 3 An aerosol solution is prepared containing the followingcomponents: Weight Active ingredient 0.25 Ethanol 25.75 Propellant 22(Chlorodifluoromethane) 74.00 Total 100.00

The active compound is mixed with ethanol and the mixture added to aportion of the propellant 22, cooled to −30° C. and transferred to afilling device. The required amount is then fed to a stainless steelcontainer and diluted with the remainder of the propellant. The valveunits are then fitted to the container.

Formulation 4 Tablets, each containing 60 mg of Active ingredient, aremade as follows: Active ingredient 60 mg Starch 45 mg Microcrystallinecellulose 35 mg Polyvinylpyrrolidone (as 10% solution in water) 4 mgSodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1 mgTotal 150 mg

The Active ingredient, starch and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The aqueous solution containingpolyvinylpyrrolidone is mixed with the resultant powder, and the mixturethen is passed through a No. 14 mesh U.S. sieve. The granules soproduced are dried at 50° C. and passed through a No. 18 mesh U.S.sieve. The sodium carboxymethyl starch, magnesium stearate and talc,previously passed through a No. 60 mesh U.S. sieve, are then added tothe granules which, after mixing, are compressed on a tablet machine toyield tablets each weighing 150 mg.

Formulation 5 Capsules, each containing 80 mg of Active ingredient, aremade as follows: Active ingredient 80 mg Starch 59 mg Microcrystallinecellulose 59 mg Magnesium stearate 2 mg Total 200 mg

The Active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 200 mg quantities.

Formulation 6 Suppositories, each containing 225 mg of Activeingredient, are made as follows: Active ingredient 225 mg Saturatedfatty acid glycerides 2,000 mg Total 2,225 mg

The Active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

Formulation 7 Suspensions, each containing 50 mg of Active ingredientper 5 ml dose, are made as follows: Active ingredient 50 mg Sodiumcarboxymethyl cellulose 50 mg Syrup 1.25 ml Benzoic acid solution 0.10ml Flavor q.v. Color q.v. Purified water to total 5 ml

The Active ingredient is passed through a No. 45 mesh U.S. sieve andmixed with the sodium carboxymethyl cellulose and syrup to form a smoothpaste. The benzoic acid solution, flavor and color are diluted with aportion of the water and added, with stirring. Sufficient water is thenadded to produce the required volume.

Formulation 8 An intravenous formulation may be prepared as follows:Active ingredient 100 mg Isotonic saline 1,000 mlThe solution of the above ingredients generally is administeredintravenously to a subject at a rate of 1 ml per minute.

The compounds of the invention are prepared by the general procedure ofreacting indole compound starting materials having carboxylfunctionality at the 4- or 5-position with an R₈₁-sulfonamide, where R₈₁is as described, supra.

All of the products of the Examples described below as well asintermediates used in the following procedures showed satisfactory nmrand ir spectra. They also had the correct elemental analyses values.

EXAMPLE A

This Example illustrates the preparation of a starting material forpreparing the acylsulfonamide functional indole compounds of theinvention (viz., formulae I and II).

Preparation of[[3-(2-amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid, a compound represented by the formula:

Part A. Preparation of 2-Ethyl-4-methoxy-1H-indole.

A solution of 140 mL (0.18 mol) of 1.3M sec-butyl lithium in cyclohexaneis added slowly to N-tert-butoxycarbonyl-3-methoxy-2-methylaniline (21.3g, 0.09 mol) in 250 mL of THF keeping the temperature below −40° C. witha dry ice-ethanol bath. The bath is removed and the temperature allowedto rise to 0° C. and then the bath replaced. After the temperature iscooled to −60° C., 18.5 g (0.18 mol) of N-methoxy-N-methylpropanamide inan equal volume of THF is added dropwise. The reaction mixture isstirred 5 minutes, the cooling bath removed and stirred an additional 18hours. It is then poured into a mixture of 300 mL of ether and 400 mL of0.5N HCl. The organic layer is separated, washed with water, brine,dried over MgSO₄, and concentrated at reduced pressure to give 25.5 g ofa crude of 1-[2-(tert-butoxycarbonylamino)-6-methoxyphenyl]-2-butanone.This material is dissolved in 250 mL of methylene chloride and 50 mL oftrifluoroacetic acid and stirred for a total of 17 hours. The mixture isconcentrated at reduced pressure and ethyl acetate and water added tothe remaining oil The ethyl acetate is separated, washed with brine,dried (MgSO₄) and concentrated. The residue is chromatographed threetimes on silica eluting with 20% EtOAc/hexane to give 13.9 g of2-ethyl-4-methoxy-1H-indole.

Analyses for C₁₁H₁₃NO:

Calculated: C, 75.40; H, 7.48; N, 7.99; Found: C, 74.41; H, 7.64; N,7.97.

Part B. Preparation of 2-Ethyl-4-methoxy-1-(phenylmethyl)-1H-indole.

2-Ethyl-4-methoxy-1H-indole (4.2 g, 24 mmol) is dissolved in 30 mL ofDMF and 960 mg (24 mmol) of 60% NaH/minerial oil is added. After 1.5hours, 2.9 mL(24 mmol) of benzyl bromide is added. After 4 hours, themixture is diluted with water and extracted twice with ethyl acetate.The combined ethyl acetate is washed with brine, dried (MgSO₄) andconcentrated at reduced pressure. The residue is chromatographed onsilica gel and eluted with 20% EtOAc/hexane to give 3.1 g (49% yield) of2-ethyl-4-methoxy-1-(phenylmethyl)-1H-indole.

Part C. Preparation of 2-Ethyl-4-hydroxy-1-(phenylmethyl)-1H-indole.

3.1 g (11.7 mmol) of 2-ethyl-4-methoxy-1-(phenylmethyl)-1H-indole isO-demethylated by treating it with 48.6 mL of 1M BBr₃/CH₂Cl₂, thenstirred at room temperature for 5 hours and concentrated at reducedpressure. The residue is dissolved in ethyl acetate, then washed withbrine and dried. The reaction product is then chromatographed on silicagel (eluted with 20% EtOAc/hexane) to give 1.58 g (54% yield) of2-ethyl-4-hydroxy-1-(phenylmethyl)-1H-indole, mp, 86-90° C.

Analyses for C₁₇H₁₇NO:

Calculated: C, 81.24; H, 6.82; N, 5.57; Found: C, 81.08; H, 6.92; N,5.41.

Part D. Preparation of[[2-Ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]acetic acid methyl ester.

2-ethyl-4-hydroxy-1-(phenylmethyl)-1H-indole (1.56 g, 6.2 mmol) istreated with 248 mg (6.2 mmol) of 60% NaH/mineral oil indimethylformamide and the mixture stirred for 0.67 hours. 0.6 mL(6.2mmol) of methyl bromoacetate is added and the mixture stirred for 17hours. The reaction mixture is diluted with water and then extractedwith ethyl acetate. The ethyl acetate solution is washed with brine,dried, and concentrated at reduced pressure. The product is purified bychromatography over silica gel eluting with 20% EtOAc/hexane, to give1.37 g (69% yield) of[[2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]acetic acid methyl ester,89-92° C.

Analyses for C₂₀H₂₁NO₃:

Calculated: C, 74.28; H, 6.55; N, 4.33; Found: C, 74.03; H, 6.49; N,4.60.

Part E. Preparation of[[3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid methyl ester.

1.36 g (4.2 mmol) of [[2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid methyl ester is reacted first with 0.4 mL (4.2 mmol) of oxalylchloride in methylene chloride and stirred for 1.5 hours. Anhydrousammonia (in excess) is bubbled into the reaction mixture and the mixtureis stirred for 1.5 hours and evaporated at reduced pressure to give awhite solid. This is stirred with ethyl acetate and the insolublematerial separated and dried to give 1.37 g of a mixture of[[3-(2-amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid methyl ester and ammonium chloride. This mixture melted at 172-187°C.

Part F. Preparation of[[3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid.

A mixture of 788 mg (2 mmol) of[3-(2-amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid methyl ester, 10 mL of 1n NaOH and 30 mL of MeOH is heated tomaintain relfux for 0.5 hour, stirred at room temperature for 0.5 hourand concentrated at reduced pressure. The residue is taken up in ethylacetate and water, the aqueous layer separated and made acidic to pH 2-3with 1N HCl. The precipitate is filtered and washed with ethyl acetateto give 559 mg (74% yield) of[[3-(2-amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl]oxy]aceticacid, mp, 230-234° C.

Analyses for C₂₁H₂₀N₂O₅:

Calculated: C, 65.96; H, 5.80; N, 7.33; Found: C, 66.95; H, 5.55; N,6.99.General Reaction Scheme used in Examples 1 to 5:

EXAMPLE 1

Preparation of1-benzyl-2-ethyl-4-methanesulfonamidoylmethyloxy-indole-3-glyoxylamide

To 100 mg (0.26 mmol) of1-benzyl-2-ethyl-4-carboxymethyloxy-indole-3-glyoxylamide suspended in3.7 ml CH₂Cl₂ was added 4-dimethylaminopyridine (48 mg, 0.39 mmol),methanesulfonamide (50 mg, 0.52 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (50 mg, 0.26mmol), respectively. After 23 h, the reaction was diluted with CH₂Cl₂and extracted with 1 N HCl, then brine. The organic layer was dried withsodium sulfate and evaporated in vacuo. The residue was chromatographedon silica gel eluting with a CH₂Cl₂/MeOH gradient to give the titledproduct (22.2 mg, 19%).

MS (ES+) 458, 413

Elemental Analyses for C₂₂H₂₃N₃O₆S:

Calculated: C 57.76; H 5.07; N 9.18 Found: C 57.66; H 5.27; N 8.92

EXAMPLE 2

Preparation of1-benzyl-2-ethyl-4-benzenesulfonamidoylmethyloxy-indole-3-glyoxylamide

To 200 mg (0.53 mmol) of1-benzyl-2-ethyl-4-carboxymethyloxy-indole-3-glyoxylamide suspended in5.3 ml CH₂Cl₂ was added 4-dimethylaminopyridine (96 mg, 0.79 mmol),benzenesulfonamide (165 mg, 1.05 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (101 mg,0.53 mmol), respectively. After 19 h, the reaction was diluted withCH₂Cl₂ and extracted with 1 N HCl, then brine. The organic layer wasdried with sodium sulfate and evaporated in vacuo. The residue waschromatographed on silica gel eluting with a CH₂Cl₂/MeOH gradient togive impure product. Recrystallation from acetone/hexane provided thetitled product (62 mg, 23%).

MS (ES+) 520, 475 Elemental Analyses for C₂₇H₂₅N₃O₆S:

Calculated: C 62.42; H 4.85; N 8.09 Found: C 62.64; H 4.97; N 8.31

EXAMPLE 3

Preparation of1-benzyl-2-ethyl-4-trifluoromethanesulfonamidoylmethyloxy-indole-3-glyoxylamide

To 200 mg (0.53 mmol) of1-benzyl-2-ethyl-4-carboxymethyloxy-indole-3-glyoxylamide suspended in5.3 ml CH₂Cl₂ was added 4-dimethylaminopyridine (96 mg, 0.79 mmol),trifluroromethanesulfonamide (157 mg, 1.05 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (101 mg,0.53 mmol), respectively. After 24 h, the reaction was diluted withCH₂Cl₂ and extracted with 1 N HCl, then brine. The organic layer wasdried with sodium sulfate and evaporated in vacuo. The residue waschromatographed on silica gel eluting with a CH₂Cl₂/MeOH gradient togive impure product. Recrystallation from acetone/hexane provided thetitled product (89 mg, 33%).

MS (ES+) 512, 467 MS (Exact)

Calculated: 512.1103 Found: 512.1108 Elemental Analyses forC₂₂H₂₀F₃N₃O₆S: Calculated: C 51.66; H 3.94; N 8.22 Found: C 49.68; H3.78; N 7.86

EXAMPLE 4

Preparation of1-benzyl-2-ethyl-4-(2-methylbenzenesulfonamidoylmethyloxy)-indole-3-glyoxylamide

To 200 mg (0.53 mmol) of1-benzyl-2-ethyl-4-carboxymethyloxy-indole-3-glyoxylamide suspended in5.3 ml CH₂Cl₂ was added 4-dimethylaminopyridine (96 mg, 0.79 mmol),2-methylbenzenesulfonamide (180 mg, 1.05 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (101 mg,0.53 mmol), respectively. After 23 h, the reaction was diluted withCH₂Cl₂ and extracted with 1 N HCl, then brine. The organic layer wasdried with sodium sulfate and evaporated in vacuo. The residue waschromatographed on silica gel eluting with a CH₂Cl₂/MeOH gradient togive impure product. Recrystallation from acetone/hexane provided thetitled product (102 mg, 36%).

MS (ES+) 534, 489 MS (Exact)

Calculated: 534.1699 Found: 534.1696 Elemental Analyses for C₂₈H₂₇N₃O₆S:Calculated: C 63.03; H 5.10; N 7.87 Found: C 63.30; H 5.87; N 7.70

EXAMPLE 5

Preparation of1-benzyl-2-ethyl-4-(4-(2-aminoethyl)benzenesulfonamidoylmethyloxy)-indole-3-glyoxylamide,Hydrochloride Salt

Part A. 4-(2-t-Butoxycarbonylamino)ethylbenzenesulfonamide

To 2.00 g (10.0 mmol) of 4-(2-aminoethyl)benzenesulfonamide suspended ina mixture of 10 ml THF, 10 ml CH₂Cl₂, and 3 ml DMF was addeddi-t-butyldicarbonate (2.20 g, 10.0 mmol) After 19 h, the reaction wasdiluted with EtOAc and extracted with 1 N HCl, then with brine. Theorganic layer was dried with sodium sulfate and evaporated in vacuo. Theresidue was triturated with hexanes and filtered to give the subtitledproduct (1.85 g, 62%).

Part B.1-benzyl-2-ethyl-4-(4-(2-t-butoxycarbonylaminoethyl)benzenesulfonamidoylmethyloxy)-indole-3-glyoxylamide

To 275 mg (0.72 mmol) of1-benzyl-2-ethyl-4-carboxymethyloxy-indole-3-glyoxylamide and 0.38 ml(2.17 mmol) N-diisopropyl,N-ethylamine suspended in 4.8 ml CH₂Cl₂ wasadded 4-dimethylaminopyridine (59 mg, 0.48 mmol),4-(2-t-butoxycarbonylamino)ethylbenzenesulfonamide (145 mg, 0.48 mmol),and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.21 g,1.09 mmol), respectively. After 24 h, the reaction was diluted withCH₂Cl₂ and extracted with 1 N HCl, then with brine. The organic layerwas dried with sodium sulfate and evaporated in vacuo. The residue waschromatographed on silica gel eluting with a hexane/acetone gradient,then with 10:1 CH₂Cl₂/MeOH to give the subtitled product (248 mg, 78%).

MS (ES+) 563, (ES−) 661 Elemental Analyses for C₃₄H₃₈N₄O₈S:

Calculated: C 61.62; H 5.78; N 8.45 Found: C 59.42; H 5.91; N 7.10

Part C.1-benzyl-2-ethyl-4-(4-(2-aminoethyl)benzenesulfonamidoylmethyloxy)-indole-3-glyoxylamide,hydrochloride salt

To 115 mg (0.17 mmol) of1-benzyl-2-ethyl-4-(4-(2-t-butoxycarbonylaminoethyl)benzenesulfonamidoylmethyloxy)-indole-3-glyoxylamidesuspended in 8.7 ml CH₂Cl₂ was added trifluoroacetic acid (4.32 ml, 56mmol). After 2 h, the reaction was evaporated in vacuo. The residue wastriturated with 5:1 CH₂Cl₂/EtOAc, filtered, and chromatographed onreverse phase silica gel eluting with an acetonitrile/water/HCl gradientto give the titled product (56 mg, 54%).

MS (ES+) 563, (ES—) 561 Elemental Analyses for C₂₉H₃₁ClN₄O₆S:

Calculated: C 58.14; H 5.22; N 9.35 Found: C 57.07; H 5.02; N 9.12

Assay

The following chromogenic assay procedure was used to identify andevaluate inhibitors of recombinant human secreted phospholipase A₂. Theassay described herein has been adapted for high volume screening using96 well microtiter plates. A general description of this assay method isfound in the article, “Analysis of Human Synovial Fluid Phospholipase A₂on Short Chain Phosphatidylcholine-Mixed Micelles: Development of aSpectrophotometric Assay Suitable for a Microtiterplate Reader”, byLaure J. Reynolds, Lori L. Hughes, and Edward A Dennis, AnalyticalBiochemistry, 204, pp. 190-197, 1992 (the disclosure of which isincorporated herein by reference):

Reagents:

REACTION BUFFER—

-   -   CaCl2.2H2O (1.47 g/L)    -   KCl (7.455 g/L)    -   Bovine Serum Albumin (fatty acid free)(1 g/L)        -   (Sigma A-7030, product of Sigma Chemical Co., St. Louis Mo.,            USA)    -   TRIS HCl (3.94 g/L)    -   pH 7.5 (adjust with NaOH)

ENZYME BUFFER—

-   -   0.05 NaOAc.3H2O, pH 4.5    -   0.2 NaCl    -   Adjust pH to 4.5 with acetic acid

DTNB—5,5′-dithiobis-2-nitrobenzoic acid

RACEMIC DIHEPTANOYL THIO—PC

racemic1,2-bis(heptanoylthio)-1,2-dideoxy-sn-glycero-3-phosphorylcholine

TRITON X-100™ prepare at 6.249 mg/ml in reaction buffer to equal 10 uM.

REACTION MIXTURE—

A measured volume of racemic dipheptanoyl thio PC supplied in chloroformat a concentration of 100 mg/ml is taken to dryness and redissolved in10 millimolar

TRITON X-100™ nonionic detergent aqueous solution. Reaction Buffer isadded to the solution, then DTNB to give the Reaction Mixture.

The reaction mixture thus obtained contains 1 mM diheptanoly thio-PCsubstrate, 0.29 mm Triton X-100™ detergent, and 0.12 mm DTMB in abuffered aqueous solution at pH 7.5.

Assay Procedure:

-   1. Add 0.2 ml reaction mixture to all wells;-   2. Add 10 ul test compound (or solvent blank) to appropriate wells,    mix 20 seconds;-   3. Add 50 nanograms of sPLA₂ (10 microliters) to appropriate wells;-   4. Incubate plate at 40° C. for 30 minutes;-   5. Read absorbance of wells at 405 nanometers with an automatic    plate reader.

All compounds were tested in triplicate. Typically, compounds weretested at a final concentration of 5 ug/ml. Compounds were consideredactive when they exhibited 40% inhibition or greater compared touninhibited control reactions when measured at 405 nanometers. Lack ofcolor development at 405 nanometers evidenced inhibition. Compoundsinitially found to be active were reassayed to confirm their activityand, if sufficiently active, IC₅₀ values were determined. Typically, theIC₅₀ values (see, Table I, below) were determined by diluting testcompound serially two-fold such that the final concentration in thereaction ranged from 45 ug/mL to 0.35 ug/ml. More potent inhibitorsrequired significantly greater dilution. In all cases, % inhibitionmeasured at 405 nanometers generated by enzyme reactions containinginhibitors relative to the uninhibited control reactions was determined.Each sample was titrated in triplicate and result values were averagedfor plotting and calculation of IC₅₀ values. IC₅₀ were determined byplotting log concentration versus inhibition values in the range from10-90% inhibition.

Results of Human Secreted Phospholipase A₂ Inhibition Tests

TABLE Compound of Inhibition of human secreted Example No. PLA₂ IC50 ±mean deviation 1 12 nM 2  7 nM 3 17 nM 4  9 nM 5 16 nMThe compounds of Example 1 to 5 are useful in inhibiting sPLA₂.

While the present invention has been illustrated above by certainspecific embodiments, it is not intended that these specific examplesshould limit the scope of the invention as described in the appendedclaims.

1. An indole compound represented by the formula (I), or apharmaceutically acceptable salt, or solvate thereof;

wherein; R₁ is selected from groups (a), (b) and (c) wherein; (a) isC₇-C₂₀ alkyl, C₇-C₂₀ haloalkyl, C₇-C₂₀ alkenyl, C₇-C₂₀ alkynyl,carbocyclic radical, or (b) is a member of (a) substituted with one ormore independently selected non-interfering substituents; or (c) is thegroup -(L₁)-R₁₁; where, -(L₁)- is a divalent linking group selected from—CH₂— or —CH₂CH₂— and R₁₁ is a group selected from (a) or (b); R₂ isselected from the group consisting of hydrogen, C₁-C₄ alkyl, C₂-C₄alkenyl, O(—C₁-C₃ alkyl), —S—(C₁-C₃ alkyl, C₃-C₄ cycloalkyl, CF₃, halo,NO₂, CN and —SO₃; R₃ is -(L₃)- Z, where -(L₃)- is a divalent linkergroup selected from a bond or a divalent group selected from:

and Z is selected from acetamide, thioacetamide, glyoxylamide,thioglyoxylamide, hydrazide or thiohydrazide groups represented by theformulae,

where R₃₁ and R₃₂ are independently selected from hydrogen, C₁-C₈ alkyl,C₁-C₈ haloalkyl, and C₃-C₄ cycloalkyl, and X is oxygen or sulfur; R₄ andR₅ each independently selected from the group consisting of hydrogen,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₇-C₁₂ aralkyl, C₇-C₁₂alkaryl, C₃-C₈ cycloalkyl, phenyl, totuyl and C₁-C₈ alkoxy or the group,-(L_(a))-(acylsulfonamide group), where -(L_(a))-, is a divalent linkerhaving a linker length of 1 to 8; provided that at least one of R₄ andR₅ must be the group, -(L_(a))-(acylsulfonamide group); and R₆ and R₇are each independently selected from the group consisting of hydrogen,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₇-C₁₂ aralkyl, C₇-C₁₂alkaryl, C₃-C₈ cycloalkyl, phenyl, totuyl and C₁-C₈ alkoxy, carbocyclicradical, carbocyclic radical substituted with C₁-C₈ alkyl, C₂-C₈ alkenyland C₇-C₁₂ alkylaryl.
 2. An indole compound represented by the formula(I), or a pharmaceutically acceptable salt, or solvate thereof;

wherein; R₁ is the group -(L₁)-R₁₁ where, -(L₁)- is —(CH₂)— and R₁₁ isC₇-C₂₀ alkyl, C₇-C₂₀ haloalkyl, C₇-C₂₀ alkenyl, C₇-C₂₀ alkynyl, or acarbocyclic radical; R₂ is —CH₃ or C₂H₅; R₃ is —C(O)—C(O)—NH₂; R₄ is thegroup -(L_(a))-(acylsulfonamide group) where -(L_(a))- is —O—CH₂— andthe acylsulfonamide group is —C(O)—NH—S(O)(O)—R₈₁ and R₈₁ is selectedfrom the group consisting of C₁-C₈ alkoxy, C₁-C₈ alkylthio, C₁-C₈alkylamino, C₂-C₈ haloalkyl, C₁-C₁₄ aralkyl, C₁-C₁₄ alkylaryl, aryl,thioaryl, C₃-C₁₄ carbocyclic radical, C₃-C₁₄ heterocyclic radical; R₅ ishydrogen; and R₆ and R₇ are hydrogen, or a carbocyclic radical.
 3. Anindole compound represented by the formula (I), or a pharmaceuticallyacceptable salt, or solvate thereof;

wherein; R₁ is the group -(L₁)-R₁₁ where, -(L₁)- is —(CH₂)— and R₁₁ isC₇-C₂₀ alkyl, C₇-C₂₀ haloalkyl, C₇-C₂₀ alkenyl, C₇-C₂₀ alkynyl, or acarbocyclic radical; R₂ is —CH₃ or C₂H₅; R₃ is —C(O)—C(O)—NH₂; R₄ is thegroup -(L_(a))-(acylsulfonamide group) where -(L_(a))- is —O—CH₂— andthe acylsulfonamide group is —C(O)—NH—S(O)(O)—R₈₁ and R₈₁ is

R₅ is hydrogen; and R₆ and R₇ are hydrogen, or a carbocyclic radical. 4.A pharmaceutical formulation comprising a indole compound as claimed inclaim 2 together with a pharmaceutically acceptable carrier or diluenttherefor.
 5. A method of treating a mammal, including a human, toalleviate the pathological effects of Inflammatory Diseases; wherein themethod comprises administration to said mammal of at least one indolecompound as claimed in claim 2 in a pharmaceutically effective amount.6. A compound selected from the group consisting of:1-benzyl-2-ethyl-4-benzezesulfonamidoylmethyloxy-indole-3-glyoxylamide;1-benzyl-2-ethyl-4-2-methylbenzezesulfonamidoylmethyloxy-indole-3-glyoxylamide;and1-benzyl-2-ethyl-4-(4-(2-aminoethyl)benzezesulfonamidoylmethyloxy-indole-3-glyoxylamidehydrochloride salt.